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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 蔣丙煌 | |
dc.contributor.author | Jia-Wen Lin | en |
dc.contributor.author | 林佳雯 | zh_TW |
dc.date.accessioned | 2021-06-15T05:43:05Z | - |
dc.date.available | 2015-08-24 | |
dc.date.copyright | 2010-08-24 | |
dc.date.issued | 2010 | |
dc.date.submitted | 2010-08-20 | |
dc.identifier.citation | 止戈。治療中草藥。五州出版社。台北,2001,p 1-5。
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/46905 | - |
dc.description.abstract | 糖尿病(diabetes mellitus;DM)為目前全球流行之慢性代謝疾病,在台灣,糖尿病多年來皆位於十大死因前五名之列,由於糖尿病患者體內長期處於高血糖之狀態,會進一步造成其病症惡化以及引發多種併發症,而飯後高血糖為造成糖尿病患許多併發症之主因之一。目前在治療糖尿病之主要藥物中,只有α-葡萄糖苷酶 (α-glucosidase) 抑制劑具有有效降低飯後血糖之效果。本實驗以抑制α-葡萄糖苷酶之活性為篩選平台,由十一種中草藥中篩選出具有抑制α-葡萄糖苷酶活性之材料,結果發現五倍子之抑制活性最為顯著,故選用五倍子做進一步的分離純化,希望從中找尋出具有調節血糖功效之活性成分,並期望能由多方面達到調節血糖以及改善糖尿病之效果。
在抑制α-葡萄糖苷酶活性試驗中,五倍子之乙酸乙酯萃取層,具有最高之抑制活性,且在抑制DPP-4酵素、抑制糖化終產物(AGEs)生成以及抑制α-澱粉酶 (α-amylase) 之實驗中,乙酸乙酯萃取層亦具有最佳之活性。進一步以 Sephadex LH-20 膠體將乙酸乙酯萃取物進行管柱層析分離為EA-A ~ EA-I九個區分層,將此九個區分層進行抑制α-葡萄糖苷酶活性試驗,實驗結果發現EA-G 區分層對酵母來源及大鼠來源之α-葡萄糖苷酶抑制性率分別為93.59 % 及42.91 %,與其他區分層相較有最佳之抑制活性。另外EA-G 區分層在清除 DPPH 自由基、抑制糖化終產物(AGEs)生成、抑制α-澱粉酶 (α-amylase) 以及促進小鼠肝細胞FL83B葡萄糖攝入之實驗中亦具有良好之活性。 由上述實驗結果選出 EA-G 區分層做進一步之純化,在經過高效液相層析儀分離純化後,得到之七個區分物G1 ~ G7,當中G4及G5兩個區分物對 α-葡萄糖苷酶之抑制活性分別為97.7 %以及99.93 %,其抑制能力顯著高於 EA-G 之其他區分物。進一步收集此兩區分物做 LC-MS 質譜分析並與文獻比對後,推知此兩個區分物為1,2,3,6-tetra-O-galloyl-β-d-glucose 及 1,2,3,4,6-penta-O-galloyl-β-d-glucose。 綜合以上實驗結果可知,五倍子具有良好之 α-葡萄糖苷酶抑制活性,並推測其主要活性成分為 1,2,3,6-tetra-O-galloyl-β-d-glucose 及 1,2,3,4,6-penta-O-galloyl -β-d-glucose。五倍子除了具有良好之 α-葡萄糖苷酶抑制活性外,尚有 α-澱粉酶抑制活性、DPP-4 抑制活性、清除 DPPH 自由基、抑制 AGEs 形成、刺激細胞吸收葡萄糖等各種生理活性,故五倍子具有由多方面性達到調節血糖之效果,以及發展為抗糖尿病活性產品之潛力。 | zh_TW |
dc.description.abstract | Diabetes mellitus (DM) is a common metabolism disease worldwide. DM is also on the top five list of the ten major causes of death in Taiwan for many years. Hyperglycemia is the main symptom to the DM patients. It obten inducesvarious complications, and postprandial hyperglycemia is the principal factor of DM complications. Among many kinds of diabetic medicines, only α-glucosidase inhibitors can significantly lower postprandial blood glucose. This research used α-glucosidase inhibition assay to find the plant materials which has ability to inhibit α-glucosidase. It was found that among the eleven plant materials tested, Galla chinensis has the most potent inhibition activity. Therefor, we futher investigated the active component in Galla chinensis usingα-glucosidase inhibition assay.
Various solvents were used to prepare Galla chinensis extracts, and ethyl acetate extract (EA) of Galla chinensis was found to have the highest ability to inhibit α-glucosidase, and it’s also had the highest activity in the DPP-4 inhibition assay, advanced glycation end products inhibition assay and α-amylase inhibition assay. We futher used Sephadex LH-20 gel to separate that EA-A ~ EA-I fractions from the Galla chinensis ethyl acetate extract, and found EA-G had the best ability to inhibit α-glucosidase among the nine fractions, the inhibition ratio of yeast α-glucosidase and rat α-glucosidase were 93.59 % and 42.91 %, respectively. The EA-G fraction also had well activity in the DPP-4 inhibition assay, DPPH free radical scavenge assay, advanced glycation end products inhibition assay and α-amylase inhibition assay, and it also increased glucose uptake of insulin resistant cells. According to the above experimental result, the EA-G fraction was futher purified using High Performance Liquid Chromatography and G1~ G7 fractions were obtained. The G4 and G5 fractions were found to have highest α-glucosidase inhibition activity among the seven fractions. These two fractions were futher analyzed by Mass Spectrometry, and found that they are 1, 2, 3, 6-tetra-O-galloyl-β-d-glucose and 1, 2, 3, 4, 6-penta-O-galloyl-β-d-glucose. This research has proved that Galla chinensis not only has high α-glucosidase inhibition activity, but also can inhibit DPP-4, scavenge free radicals, inhibit advanced glycation end products formation, inhibit α-amylase activity, and increase glucose uptake of insulin resistant cells. So, Galla chinensis can modulate blood sugar through various ways and has potential to be developed as an anti-diabetes product. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T05:43:05Z (GMT). No. of bitstreams: 1 ntu-99-R97641019-1.pdf: 2765461 bytes, checksum: 5bc1159a38f77fda85a05a8474ed18b5 (MD5) Previous issue date: 2010 | en |
dc.description.tableofcontents | 第一章 前言 ………………………………………………………………………………………………… 1
第二章 文獻回顧 ..………………………………………………………………………………………………. 3 第ㄧ節 糖尿病 ………………………………………………………………………………………………. 3 一、糖尿病基本介紹 ..…………………………………………………………………………………. 3 二、糖尿病診斷依據及分類 .…………………………………………….….……………..……… 3 (一)糖尿病診斷依據 ……………….….……………………………..…………..…………… 3 (二)糖尿病分類 …..………………………………………………………………..……..……… 4 1. 第一型糖尿病 …………………………………….………………………..….…………… 4 2. 第二型糖尿病 ……………………………………………………………..…..…………… 4 3. 妊娠糖尿病 …………………………………………………………………….…………… 5 4. 其他特殊因素糖尿病 ………………………………………………………….………… 5 三、第二型糖尿病致病機制與高血糖對人體之影響 ……………….……….………… 6 (一)第二型糖尿病之致病機制 …………………………….………………….…….….…… 6 (二)高血糖對人體之影響 …………………….………………………………….……..……… 7 1. 高血糖與糖尿病併發症 ……………….…………….……….……………………..… 7 2. 高血糖與氧化壓力 ….………………….……………….…………………..…………… 7 3. 高血糖與糖化終產物 …………………...…………….………………………..…….… 9 四、糖尿病治療與用藥 …………………………………………………………………….…..……… 11 (一) 雙胍類藥物(Biguanides) …………………...…………….……………………….....… 11 (二) Thiazolidinedione 類藥物 ……………...…………….…………………….……….…. 11 (三) α-glucosidase抑制劑 ………….……………………………………………….…....………12 (四) 磺醯尿素類藥物(sulfonylurea, SU) ..……………….………………………....……12 (五) 非磺醯尿素類藥物(Non-Sulfonylurea) ………..….………………….……....……13 (六) Glucagon-like peptide 1(GLP-1) 類似物 ……..….……………………….....……13 (七) Dipeptidyl peptidase-4 (DPP-4) 抑制劑 ……….….…………………………..……14 第二節α-葡萄糖苷酶抑制劑 (α-Glucosidase inhibitor) ..……..…………………….…… 15 一、α-葡萄糖苷酶 (α-Glucosidase)介紹 ..………………………………….………….……... 15 二、α-葡萄糖苷酶 (α-Glucosidase) 抑制劑 …………….………………….………..…….. 16 三、α-葡萄糖苷酶 (α-Glucosidase) 抑制劑於第二型糖尿病中之作用 ……....…17 四、α-葡萄糖苷酶 (α-Glucosidase) 抑制劑於天然物中之研究 ………..……...…. 21 第三節 五倍子之研究回顧 ………………………..……….……………………..……………….…..… 24 一、五倍子介紹 ………...…………………………………………………………..……………..…..….. 24 二、五倍子主要化學成分 ..…………………………………………………………..................... 25 三、五倍子生理活性之研究 ..………………………………………………………………….….…. 28 四、五倍子於治療糖尿病之研究 ………………………………………..……………………..…. 30 第三章 研究目的與實驗架構 …………………………………………………..………………………….… 31 一、研究目的 ………........…………………………………………………………..………………….…. 31 二、實驗架構 ………........…………………………………………………………..…………………….. 32 (一) 以抑制 α-glucosidase 為篩選平台,由多種中草藥中篩選出具有 抑制 α-glucosidase 潛力之材料 ……………………………..……………………... 32 (二) 探討五倍子經不同溶劑進行萃取後抑制 α-glucosidase 之能力 …... 33 (三) 五倍子乙酸乙酯萃取物之分離純化 ……………………..………………….…….. 34 (四) 五倍子乙酸乙酯萃取物之各區分層對抗糖尿病活性試驗 …..…….….. 35 (五) 五倍子乙酸乙酯萃取物之各區分層成分分析鑑定 …………………….….. 36 第四章 實驗材料與方法 ……………………………………………………………………………………..…. 37 一、中藥材來源 ………………………………………………………………………..………………….… 37 二、實驗方法與儀器藥品 ………………………………………………………………………….…… 37 (一) 五倍子水萃物製備 …………………………………………………………………..…..…. 37 (二) 五倍子液相萃取實驗 …………………………………………………………………..….. 37 (三) α- Glucosidase 抑制實驗 …..……………………………………….…………………..….38 (四) 五倍子乙酸乙酯萃取物管柱層析實驗 …..……………………………….…..…. 39 (五) 總酚含量測定實驗 …………………………………………………………………….….…. 40 (六) 總類黃酮含量測定 …..………………………………………………………………...……. 41 (七) DPP-4酵素抑制實驗 ……………………………………………..…………………..……. 42 (八) 糖化終產物抑制實驗 …..…………………………………………………………….….…. 43 (九) DPPH自由基清除實驗 …………………………………………………….………..……. 45 (十) 促進細胞攝入葡萄糖 (Glucose uptake) 實驗 ………………..………………. 46 (十一) α- Amylase 抑制實驗 …………………………………………….……..…..…..……. 50 (十二) 高效液相層析儀分析實驗 …………………….……………….……..…………. 51 (十三) 電噴灑游離質譜分析 …………………….………………..…….……..…………. 51 (十四) 數據分析 …………………….………………………….………..…….……..…………. 52 第五章 結果與討論 ……………………………………………………………………………..……………... 53 一、由多種中草藥中篩選出具有α- Glucosidase 抑制活性之材料 ……….…..54 二、五倍子經不同溶劑萃取後之活性試驗.……………………………………..………… 56 (一) 五倍子水萃物經不同溶劑萃取之回收率………………………………..…… 56 (二) 五倍子水萃物經不同溶劑萃取後對 α-glucosidase 之抑制能力… 57 (三) 五倍子水萃物經不同溶劑萃取後對dipeptidyl peptidase-IV( DPP-4) 酵素之抑制能力………………..……………………………………………..……..…... 60 (四) 五倍子水萃物經不同溶劑萃取後抑制糖化終產物 (AGEs) 形成之 能力…………………………………………………………………………..……….….……... 62 (五) 五倍子水萃物經不同溶劑萃取後抑制澱粉水解酶(α-amylase) 之 能力…………………………………………………………………………..……….….……... 62 (六) 小結…………………………………………………………………………..……….….……... 65 三、五倍子水萃物乙酸乙酯萃取物之管柱分離…………………..…..……….….…….. 66 (一) 經 Sephadex LH-20 膠體分離後之薄層層析結.………………………… 66 (二) 各區分層之回收率 ………….………………………………………………………….. 66 四、五倍子水萃物乙酸乙酯萃取物各區分層之活性試驗.……………………….... 70 (一) 五倍子水萃物乙酸乙酯萃取物各區分層對α-glucosidase 之抑制能 力……………………………………………………………………………………………..…… 70 (二) 五倍子水萃物之乙酸乙酯萃取物各區分層之總酚及總類黃酮含 量……………………………………………………………………………………………………73 (三) 五倍子水萃物之乙酸乙酯萃取物各區分層對dipeptidyl peptidase-IV ( DPP-4) 酵素之抑制能力…………………………………………….……..…... 77 (四) 五倍子水萃物之乙酸乙酯萃取物各區分層抑制糖化終產物 (AGEs) 形成之能力…………………..……………………………………………………………... 79 (五) 五倍子水萃物之乙酸乙酯萃取物各區分物清除DPPH自由基之能 力………………………………………………………………………..……….……….……... 81 (六) 五倍子水萃物之乙酸乙酯萃取物各區分層抑制澱粉水解酶 (α-amylase) 之能力…….…………………………………………………………………………….……... 83 (七) 五倍子水萃物之乙酸乙酯萃取物各區分層促進小鼠肝細胞 FL83B 葡萄糖攝入之能力.……………………………………………………………………… 85 (八) 小結……………………………………………………………………………………………… 87 四、五倍子水萃物之乙酸乙酯萃取物EA-G區分層之活性成分分析…….… 89 (一) EA-G 區分層之 HPLC 分析………………………………………………………89 (二) EA-G 區分層之各區分物對於 α-葡萄糖苷酶 之抑制能力……....89 (三) G4 與 G5 區分物之質譜儀分析………………………………………….…....89 第六章 綜合討論………………………………………………………………………..……………………….. 96 第七章 結論 ………………………………………………………………………..…………………………….. 98 第八章 參考文獻 …………………………………………………………………………………………….…. 99 | |
dc.language.iso | zh-TW | |
dc.title | 以α-葡萄糖苷酶之活性抑制為主軸探討五倍子之調節血糖功能與成分 | zh_TW |
dc.title | Anti-hyperglycemia activity of Galla chinensis based on the inhibition of α- glucosidase activity | en |
dc.type | Thesis | |
dc.date.schoolyear | 98-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 孫璐西,何其儻,游丞德 | |
dc.subject.keyword | 糖尿病,調節血糖,五倍子,α-葡萄糖苷,酶,抑制劑, | zh_TW |
dc.subject.keyword | diabetes mellitus,Galla chinensis,α-glucosidase inhibitor,1 ,2,3,6-tetra- O-galloyl-β-d-glucose,1,2,3,4,6-penta-O-galloyl-β-d-glucose, | en |
dc.relation.page | 112 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2010-08-20 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 食品科技研究所 | zh_TW |
顯示於系所單位: | 食品科技研究所 |
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